Selection of antigen retrieval techniques is crucial for successful staining of paraffin sections. Different antibodies may require different antigen retrieval techniques and need to be tested prior to application of actual projects.

E.Primary Antibody:Dilution of primary antibody is critical for success of staining, so a titration must be performed prior to application of actual projects.

F.Secondary Antibody:An appropriate biotinylated secondary antibody should be selected and a titration is needed prior to application of actual projects.

G.ABC Reagent:Depending on sensitivity and morphological requirements, there are varieties of ABC kits and reagents can be selected from. The most commonly used one is VECTASTAIN ABC Kit from Vector Laboratories.

H.AEC Reagent

Procedure

1.Deparaffinize sections in xylene for 3x5min.2.Hydrate with 100% ethanol for 2x5min.3.Hydrate with 95% ethanol for 2x5min.4.Rinse in distilled water.

5.Antigen Retrieval: (Select an appropriate antigen retrieval technique and it depends on antibodies used).6.Rinse sections in PBS for 1x5min.